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    Glucose Determination

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    Via Gail Canlas
    1. This document discusses glucose determination through quantitative enzymatic-colorimetric methods. 2. It involves using glucose oxidase, peroxidase, and phenol to produce a quinone complex from hydrogen peroxide and 4-aminoantipyrine, whose absorbance is used to calculate glucose concentration in a sample. 3. The pre-analytical phase requires plasma collection within 30 minutes using fluoride and the analytical phase involves incubating and reading samples and standards to obtain absorbance values for calculation.

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    Glucose Determination

    Uploaded by

    Via Gail Canlas
    31 views12 pages
    1. This document discusses glucose determination through quantitative enzymatic-colorimetric methods. 2. It involves using glucose oxidase, peroxidase, and phenol to produce a quinone complex from hydrogen peroxide and 4-aminoantipyrine, whose absorbance is used to calculate glucose concentration in a sample. 3. The pre-analytical phase requires plasma collection within 30 minutes using fluoride and the analytical phase involves incubating and reading samples and standards to obtain absorbance values for calculation.

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    1. This document discusses glucose determination through quantitative enzymatic-colorimetric methods. 2. It involves using glucose oxidase, peroxidase, and phenol to produce a quinone complex from hydrogen peroxide and 4-aminoantipyrine, whose absorbance is used to calculate glucose concentration in a sample. 3. The pre-analytical phase requires plasma collection within 30 minutes using fluoride and the analytical phase involves incubating and reading samples and standards to obtain absorbance values for calculation.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    31 views12 pages

    Glucose Determination

    Uploaded by

    Via Gail Canlas
    1. This document discusses glucose determination through quantitative enzymatic-colorimetric methods. 2. It involves using glucose oxidase, peroxidase, and phenol to produce a quinone complex from hydrogen peroxide and 4-aminoantipyrine, whose absorbance is used to calculate glucose concentration in a sample. 3. The pre-analytical phase requires plasma collection within 30 minutes using fluoride and the analytical phase involves incubating and reading samples and standards to obtain absorbance values for calculation.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 12

    GLUCOSE

    DETERMINATION

    KIMBERLY ANN M. PULGA, RMT, MPH


    COLLEGE OF MEDICAL LABORATORY SCIENCE
    OUR LADY OF FATIMA UNIVERSITY
    VALENZUELA CAMPUS
    UNIT OUTCOMES

    To achieve this unit a learner must:

    1. Rationalize the consideration in patient preparation, specimen


    collection, transport and processing and handling.
    2. Describe the specimen of choice, principle involved, advantages
    of the different laboratory methods of glucose determination.
    3. Be able to properly perform fasting blood sugar determination.
    4. Be able to evaluate FBS results correctly.
    PRE-ANALYTICAL PHASE

    Method: Quantitative Enzymatic-Colorimetric Determination of


    Glucose in Serum or Plasma

    Principle:
    PRE-ANALYTICAL PHASE

    Method: Quantitative Enzymatic-Colorimetric Determination of


    Glucose in Serum or Plasma

    Principle:
    GO
    Glucose + O2 + H2O à Gluconic acid + H2O2

    PO
    2H2O2 + 4-aminoantipyrine + Phenol à Quinone complex
    PRE-ANALYTICAL PHASE

    Specimen Collection and Preparation:

    Remove from the clot within 30 minutes of collection in order to


    prevent glycolysis.

    Plasma: an anticoagulant containing fluoride is recommended.


    ANALYTICAL PHASE

    Materials:
    Spectrophotometer
    Accurate pipetting devices
    Heating block or water bath (100 degrees Celsius)
    Cuvettes
    Vortex mixer
    Interval timer
    ANALYTICAL PHASE

    Reagents:
    Glucose liquicolor reagent Non-reactive ingredients & preservatives
    4-aminoantipyrine preservatives
    Glucose oxidase Glucose standard (100 mg/dL)
    Peroxidase Glucose in aqueous benzoic acid
    Phenol

    Storage and stability: Stable at 2-8 degrees Celsius


    Do not freeze
    ANALYTICAL PHASE

    Manual Procedure:
    1. Pipet into tubes the following REAGENT STANDARD (S) SAMPLE (U)
    volumes (mL) and mix well: BLANK (RB)

    REAGENT 1.5 1.5 1.5


    2. Incubate all tubes at 100
    degrees Celsius for 3 minutes. STANDARD - 0.050 -

    SAMPLE - - 0.050

    3. Immerse test tubes in room


    temperature water for 2
    minutes to cool. Read S and U
    vs. RB at 600 nm.
    ANALYTICAL PHASE

    Glucose Determination

    Glucose Determination 2
    POST-ANALYTICAL PHASE

    Formula for glucose concentration:

    C (mg/dL) = __Abs sample___ x value of standard (100mg/dL)


    Abs standard

    C (mmol/L) = __Abs sample___ x value of standard (100mg/dL) x 0.056


    Abs standard
    POST-ANALYTICAL PHASE

    Normal values (Profame):


    Serum/Plasma (fasting): 65-110mg/dL
    Spinal fluid: 40-70 mg/dL

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